An indirect immunoassay for quantitative determination of ampicillin (range, 10–1000 ng/ml) in buffer or milk has been developed. Polyclonal antibodies were obtained against ampicillin conjugated with bovine serum albumin; the conjugate was synthesized by direct condensation using carbodiimide. The antibodies were specific for ampicillin and exhibited low cross-reactivity to other penicillins (azlocillin, 17%; penicillin G, 10%; piperacillin, 5%; and carbenicillin, 4%). Matrix effects were minimized by combining the use of a casein-supplemented buffer (content of casein, 1%) with sample dilution. Limit of detection for ampicillin in milk (diluted tenfold) was equal to 5.0 ng/ml (which corresponded to 50 ng/ml of the original sample).

An indirect immunoassay for quantitative determination of ampicillin (range, 10-1000 ng/ml) in buffer or milk has been developed. Polyclonal antibodies were obtained against ampicillin conjugated with bovine serum albumin; the conjugate was synthesized by direct condensation using carbodiimide. The antibodies were specific for ampicillin and exhibited low cross-reactivity to other penicillins (azlocillin, 17%; penicillin G, 10%; piperacillin, 5%; and carbenicillin, 4%). Matrix effects were minimized by combining the use of a casein-supplemented buffer (content of casein, 1%) with sample dilution. The threshold of ampicillin detection in milk (diluted tenfold) was equal to 5.0 ng/ml (which corresponded to 50 ng/ml of the original sample).

The method of solid-phase enzyme linked immunosorbent assay (ELISA) for quantitative detection of chloramphenicol (CAP) in milk was developed. Peculiarities of the adsorption on the microtitre plates surface of CAP-ovalbumin conjugate were investigated. Different conditions of competition stage of the analysis were studied. Conditions providing CAP monitoring in human blood serum in the clinical range were optimized. Matrix effect on the assay results was studied. The specificity of the analytical system was investigated and the reagents stability was examined. The method developed permits CAP concentration to be determined in human blood serum, diluted 1/100, in the linear range from 0.1 to 100 ng/ml. The assay is characterized by high sensitivity (0.08 ng/ml) and good reproducibility (CV < 10.8%), assay time is about 3 hours. The correlation coefficient with HPLC is 0.977.

Оглавление: 1. Ферменты 1.1. Щелочная фосфатаза 1.2. Аминотрансферазы 1.3. α-Амилаза 1.4. Лактатдегидрогеназа 1.5. Креатинкиназа 1.6. γ-Глутамилтрансфераза 2. Субстраты и белки 2.1. Мочевина 2.2. Креатинин 2.3. Мочевая кислота 2.4. Белок общий 2.5. Альбумин 2.6. Билирубин 2.7. Глюкоза 2.8. Холестерин и фракции холестерина 2.9. Триглицериды 2.10. Железо, ЛЖСС, ОЖСС 2.11. Кальций 2.12. Магний 2.13. Калий 2.14. Натрий 2.15. Хлориды 2.16. Фосфор неорганический 2.17. Гликозилированный гемоглобин 2.18. С-реактивный белок 3. Правила подготовки пациента для проведения биохимических исследований 4. Литература

An enzyme immune test system was designed and optimized for quantitative assay of gentamicin in human sera. Immunospecific reagents i.e. gentamicin conjugates with ovalbumin (for sorption on polysterol plates) and with bovine serum albumin (immunogen) were prepared. Gentamicin specific antisera were isolated and tested. Conditions for the antigen sorption on polysterol plates were determined and optimized. Different regimes of the competition reaction were investigated and conditions for the antibiotic assay in human sera were determined. The assay specificity was studied and the stability of the test system was checked. An experimental lot of the reagent set was manufactured at the ZAO NPP Immunotech and the correlation tests with the use of the fluorescence polarization immunoassay were performed. The set is destined for the assay of 40 samples (in duplicate). The method sensitivity is 1 ng/ml of gentamicin. The range of the detectable concentration is 1 to 32 ng/ml of gentamicin in 1000-fold diluted sera. The assay time is not more than 3 hours. The variation coefficient of the results does not exceed 12 per cent. The shelf-life of the set is 0.5 years when stored at a temperature of 2 to 8 degrees C.

The method of enzyme linked immunosorbent assay (ELISA) for quantitative detection of chloramphenicol (CAP) in human blood serum was developed. Peculiarities of the adsorption on the microtitre plates surface of CAP-ovalbumin conjugate were investigated. Different conditions of competition stage of the analysis were studied. Conditions providing CAP monitoring in human blood serum in the clinical range were optimized. Matrix effect on the assay results was studied. The specificity of the analytical system was investigated and the reagents stability was examined. The method developed permits CAP concentration to be determined in human blood serum, diluted 1/100, in the linear range from 10 to 1000 ng/ml. The assay is characterized by high sensitivity (1 ng/mL) and good reproducibility (CV < 12%), assay time is about 3 hours.

Polarization fluoroimmunoassay for s-triazine herbicides was developed. The sensitive of assay is higher with using shortest chemical "bridge" length between the molecule of antigen and fluorescent label. The detection limit of atrazine in 50 microL sample was 6 ng/ml. The total time required for an assay of 10 samples was 6 ng/ml. The total time required for an assay of 10 samples is approximately 7 min. The method may be used for environmental monitoring of s-triazine herbicides in water samples.

The review deals with studies published during the last 10 years on the development of immunochemical methods for the assay of herbicides of the 1,3,5-triazine group. Questions concerning the optimisation of methods for the immunoenzyme assay and their application in the determination of 1,3,5- triazines in various samples (water, soil, food products etc.) are discussed. The bibliography includes 75 references.

Influence of labelled antigens structure on the sensitivity of the polarization fluoroimmunoassay of atrazine was studied. It is shown that the highest sensitivity is provided by the use of the heterologous labelled reagent with the shortest chemical bridge between the antigen and fluorescent label.